Antitumor Effect And Mechanisms Of Capilliposide Hydrolysate LC-A From Lysimachia Capillipes Hemsl

Objective: Preparatory work showed capilliposide(LC) from Lysimachia capillipes Hemsl has significant anti-tumor effects in vivo, but also in vitro. Firstly, the effect in vitro proliferation of several human tumor cell lines inhibition was evaluted, then statistical analysis was carried out to obtain the anti-tumor spectrum of LC-A. To observe the inhibitory effect on the growth of transplanted tumor in nude mice and evaluate the in vivo efficacy, the nude mice xenografts were established with the most sensitive tumor cells selected by in vitro screening; determine the in vitro and in vivo pharmacodynamics preliminary the mechanism were investigated.Method: In vitro, a variety of anthropogenic tumor cell lines such as prostate cancer PC-3, liver cancer HepG2, gastric cancer BGC823, lung cancer A549, colon cancer SW620, breast cancer MDA-MB-231 as the research object, the cells were exposed to LC-A in a series of concentration gradient(20, 10, 5, 2.5, 1.25μΜ/L) for 48 h, OD value was measured by enzyme labeled instrument. The effect in vitro proliferation of several human tumor cell lines inhibition of capilliposide hydrolysate LC-A from Lysimachia capillipes Hemsl was evaluted determined by MTT method, and calculated the cell proliferation inhibition rate of every drug concentration, then analysis the datas and calculated the half inhibitory concentration IC50 by SPSS17.0.In vivo, people colon cancer SW620 xenograft in nude mice tumor model was established, adjust the cells to a certain concentration, and inoculated to the axillary region of mice. Selected mice were randomly grouped and administration in oral everyday for 4 weeks, then mice were sacrificed and dissected tumor tissue, calculated the tumor inhibition rate, relative tumor growth rate(T/C) and the index of thymus and spleen, evaluated the inhibitory effect on tumor.About the mechanisms, SW620 cells were cultured in different concentrations(10, 5 and 2.5μΜ/L) of LC-A for 48 h,then cells were collected, total protein was extracted of cells and tumor for Western blot experiments, to detect the antitumor molecular mechanisms.Results: LC-A has certain inhibitory effect in PC-3 prostate cancer, hepatocellular carcinoma HepG2, gastric cancer BGC823, lung cancer A549, colon cancer SW620 and breast cancer MDA-MB-231 cells, with IC50 values were 13.44, 25.72, 8.40, 11.19, 5.93, and 13.33μΜ/L, thus LC-A has broad-spectrum antitumor properties, especially for colon cancer.In vivo, compared with the control group, the growth rate of the tumor bearing nude mice was slowed down after administration, and with the increase of the LC-A drug dose, the tumor growth rate was decreased, with positive correlation with drug dosage. The growth of human colorectal cancer SW620 tumor of oral administration group(50 and 100 mg/kg) and intraperitoneal injection group(15 mg/kg) were inhibited in different extent, with tumor inhibition rate were 24.41%, 63.05% and 43.61%, positive control group CTX tumor inhibition rate was 54.87%.About the mechanism, LC-A upgraded the expression of LC-3B protein, reduce the phosphorylation level of PI3 K, Akt and mTor, and interfere with the cell signal transduction pathway of PI3K/Akt/mTor,it May also lead to excessive activation of PARP.Conclusion: Capilliposide hydrolysate LC-A from Lysimachia capillipes Hemsl has significant antitumor effect in vitro and in vivo. Western blot suggested that LC-A could up-regulate the expression of LC-3B and induced autophagy by intervention the cell signal transduction pathway of PI3K/Akt/mTor, Non-Caspase dependent apoptosis can be induced by excessive activation of PARP in vitro.